TABLE 1.
Primers used in this study
| Primer purpose and primer | Sequencea |
|---|---|
| Disruption primers for hyg19 gene | |
| Forward | GATCCGACCCGTGCCGGGCGGAAGGGAGCACCAGTGATGATTCCGGGGATCCGTCGACC |
| Reverse | GGTGATGAGCGGGGCGCTGAGCCAGAGGAACATCACGACTGTAGGCTGGAGCTGCTTC |
| Disruption primers for hyg28 gene | |
| Forward | CTGATGACTGCCACTCTCGTCGCCAAGGATCTGGCCGCCATTCCGGGGATCCGTCGACC |
| Reverse | CTCGGTGACCCGGCCGTCCGCCACCTCCAGGCGGCGGGTTGTAGGCTGGAGCTGCTTC |
| Disruption primers for hyg19, hyg20, and hyg21 genes | |
| Forward | GATCCGACCCGTGCCGGGCGGAAGGGAGCACCAGTGATGATTCCGGGGATCCGTCGACC |
| Reverse | ACCGTTGACCGAGAATGGGTAAAGGAGCAGAAAACAATGTGTAGGCTGGAGCTGCTTC |
| Expression primers for hyg26 gene | |
| Forward | CATATGAGTGGACTGATGCGGGAC |
| Reverse | GGATCCTCAGAATTCGCTGACGC |
a
The 39-nucleotide homologous region flanking the targeted gene is indicated in boldface. The italicized primer region is homologous to pIJ773. The NdeI and BamHI sites in hyg26 expression primers are also indicated in boldface.